| Assay Method Information | |
| | RNAse H Inhibition Assay |
| Description: | To compare activity and inhibition of E. coli RNase H with HIV-1 RNase H, theconcentration of the E. coli was adjusted to the efficiency of 500 nM HIV-1 RT in the absence of inhibitor. For IC50 determination, 100 nM of 5'-radiolabeled PBS-14r8d was heat annealed to a 3-fold molar excess as described above. 100 nM preformed DNA-RNA/DNA hybrid was incubated with 500 nM HIV-1 RT or 10 nM E. coli RNase Hin a buffer of 50mM Tris-HCl (pH 7.8), 50 mM NaCl, 0.2 mM EDTA, and varying concentrations of GSK5750 and β-thujaplicinol. Reactions were initiated by theaddition of 6 mM MgCl2 and allowed to proceed at 37 °C for 6 min. Samples were analyzed as described above. |
| Affinity data for this assay | |
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